Elucidation of structure-function relationship of THCA and CBDA synthase from Cannabis sativa L.

Journal of Biotechnology

“Cannabinoids are secondary natural products from the plant Cannabis sativa L.

Therapeutic indications of cannabinoids currently comprise a significant area of medicinal research.

We have expressed the Δ9-tetrahydrocannabinolic acid synthase (THCAS) and cannabidiolic acid synthase (CBDAS) recombinantly in Komagataella phaffii and could detect eight different products with a cannabinoid scaffold after conversion of the precursor cannabigerolic acid (CBGA).

Besides five products remaining to be identified, both enzymes were forming three major cannabinoids of C. sativa – Δ9-tetrahydrocannabinolic acid (THCA), cannabidiolic acid (CBDA) and cannabichromenic acid (CBCA).

These studies lay the groundwork for further research as well as biotechnological cannabinoid production.”

Facebook Twitter Pinterest Stumbleupon Tumblr Posterous

Anti-Inflammatory Activity in Colon Models Is Derived from Δ9-Tetrahydrocannabinolic Acid That Interacts with Additional Compounds in Cannabis Extracts.

“Inflammatory bowel diseases (IBDs) include Crohn’s disease, and ulcerative colitis. Cannabis sativa preparations have beneficial effects for IBD patients. However, C. sativa extracts contain hundreds of compounds. Although there is much knowledge of the activity of different cannabinoids and their receptor agonists or antagonists, the cytotoxic and anti-inflammatory activity of whole C. sativa extracts has never been characterized in detail with in vitro and ex vivo colon models.

Material and Methods: The anti-inflammatory activity of C. sativa extracts was studied on three lines of epithelial cells and on colon tissue. C. sativa flowers were extracted with ethanol, enzyme-linked immunosorbent assay was used to determine the level of interleukin-8 in colon cells and tissue biopsies, chemical analysis was performed using high-performance liquid chromatography, mass spectrometry and nuclear magnetic resonance and gene expression was determined by quantitative real-time PCR.

Results: The anti-inflammatory activity of Cannabis extracts derives from D9-tetrahydrocannabinolic acid (THCA) present in fraction 7 (F7) of the extract. However, all fractions of C. sativa at a certain combination of concentrations have a significant increased cytotoxic activity. GPR55 receptor antagonist significantly reduces the anti-inflammatory activity of F7, whereas cannabinoid type 2 receptor antagonist significantly increases HCT116 cell proliferation. Also, cannabidiol (CBD) shows dose dependent cytotoxic activity, whereas anti-inflammatory activity was found only for the low concentration of CBD, and in a bell-shaped rather than dose-dependent manner. Activity of the extract and active fraction was verified on colon tissues taken from IBD patients, and was shown to suppress cyclooxygenase-2 (COX2) and metalloproteinase-9 (MMP9) gene expression in both cell culture and colon tissue.

Conclusions: It is suggested that the anti-inflammatory activity of Cannabis extracts on colon epithelial cells derives from a fraction of the extract that contains THCA, and is mediated, at least partially, via GPR55 receptor. The cytotoxic activity of the C. sativa extract was increased by combining all fractions at a certain combination of concentrations and was partially affected by CB2 receptor antagonist that increased cell proliferation. It is suggested that in a nonpsychoactive treatment for IBD, THCA should be used rather than CBD.”

Facebook Twitter Pinterest Stumbleupon Tumblr Posterous

Cannabis tea revisited: a systematic evaluation of the cannabinoid composition of cannabis tea.

“Cannabis is one of the oldest known medicinal plants, and a large variety of biological activities have been described. The main constituents, the cannabinoids, are thought to be most important for these activities. Although smoking of cannabis is by far the most common way of consumption, a significant part of medicinal users consume it in the form of a tea.

However, not much is known about the composition of cannabis tea, or the effect of different parameters during preparation, handling or storage. In this study we used the high-grade cannabis available in Dutch pharmacies to study the cannabinoid composition of tea under standardized and quantitative conditions. Experimental conditions were systematically varied in order to mimic the possible variations made by medicinal users.

During analysis there was a specific focus on the cannabinoid tetrahydrocannabinol and its acidic precursor, tetrahydrocannabinolic acid. Also the role of non-psychoactive cannabinoids as components of cannabis tea are discussed.

The results obtained in this study provide a clear quantitative insight in the phytochemistry of cannabis tea preparation and can contribute to a better appreciation of this mode of cannabis administration.”

http://www.ncbi.nlm.nih.gov/pubmed/17604926 

Facebook Twitter Pinterest Stumbleupon Tumblr Posterous